The influence of prostacyclin (PGI2) on contractile properties of isolated right ventricle of rat heart

The mechanism of the positive inotropic effect of prostacyclin (PGI₂) (2.6×10^?6 mol/l) on the isolated right ventricle of rat heart was studied. Our results show that the positive inotropic effect of prostacyclin is produced indirectly through beta adrenoceptors and slow Ca²⁺ channels, because blockade of slow Ca²⁺ channels with verapamil (10^?6 mol/l) and beta adrenoceptors with propranolol (10^?6 mol/l) abolishes this effect. Alpha adrenoceptors do not mediate the action of PGI₂.     

Increase in in vivo (3H) spiperone binding in the rat hippocampal formation and striatum after repeated treatment with haloperidol

Summary An increase in in vivo (³H) spiperone binding was observed in rat hippocampal formation and striatum after repeated treatment with haloperidol. This suggests that in hippocampus as well as in striatum prolonged blockade of dopaminergic transmission by a neuroleptic agent results in the development of a supersensitivity of the dopamine receptors.Acknowledgments. We wish to thank Mrs J. Krauss for skillful technical assistance, Drs Helmut Bittiger and Rainer Ortmann for helpful discussions.     

Muscarinic involvement in vascular and adrenal medullary responses to splanchnic nerve stimulation in conscious calves

Stimulation of the peripheral end of the right splanchnic nerve (4 Hz for 10 min) in the presence of hexamethonium caused a small but significant rise in mean aortic blood pressure which was subsequently abolished by atropine. There were also small but significant increases in the outputs of catecholamines, [Met⁵]-enkephalins and corticotrophin releasing factor (CRF) from the right adrenal gland. The catecholamine response was roughly halved after atropine while the outputs of enkephalins and CRF were unaffected. It is concluded that splanchnic sympathetic postganglionic neurones supplying the vasculature are completely blocked by cholinergic blockade whereas adrenal medullary responses persist in an attenuated form.     

Complex modulation of Cav3.1 T-type calcium channel by nickel

Nickel is considered to be a selective blocker of low-voltage-activated T-type calcium channel. Recently, the Ni²⁺-binding site with critical histidine-191 (H191) within the extracellular IS3–IS4 domain of the most Ni²⁺-sensitive Ca_v3.2 T-channel isoform has been identified. All calcium channels are postulated to also have intrapore-binding site limiting maximal current carried by permeating divalent cations (PDC) and determining the blockade by non-permeating ones. However, the contribution of the two sites to the overall Ni²⁺ effect and its dependence on PDC remain uncertain. Here we compared Ni²⁺ action on the wild-type “Ni²⁺-insensitive” Ca_v3.1_w/t channel and Ca_v3.1_Q172H mutant having glutamine (Q) equivalent to H191 of Ca_v3.2 replaced by histidine. Each channel was expressed in Xenopus oocytes, and Ni²⁺ blockade of Ca²⁺, Sr²⁺, or Ba²⁺ currents was assessed by electrophysiology. Inhibition of Ca_v3.1_w/t by Ni²⁺ conformed to two sites binding. Ni²⁺ binding with high-affinity site (IC₅₀ = 0.03–3 μM depending on PDC) produced maximal inhibition of 20–30 % and was voltage-dependent, consistent with its location within the channel’s pore. Most of the inhibition (70–80 %) was produced by Ni²⁺ binding with low-affinity site (IC₅₀ = 240–700 μM). Q172H-mutation mainly affected low-affinity binding (IC₅₀ = 120–160 μM). The IC₅₀ of Ni²⁺ binding with both sites in the Ca_v3.1_w/t and Ca_v3.1_Q172H was differentially modulated by PDC, suggesting a varying degree of competition of Ca²⁺, Sr²⁺, or Ba²⁺ with Ni²⁺. We conclude that differential Ni²⁺-sensitivity of T-channel subtypes is determined only by H-containing external binding sites, which, in the absence of Ni²⁺, may be occupied by PDC, influencing in turn the channel’s permeation.     

In vitro dopaminergic neuroprotective and in vivo antiparkinsonian-like effects of Δ3,2-hydroxybakuchiol isolated from Psoralea corylifolia (L.)

Cocktail recipes containing Psoralea corylifolia seeds (PCS) are used to empirically treat Parkinson disease. A PCS isolate Δ³,2-hydroxybakuchiol (BU) can inhibit dopamine uptake in dopamine transporter (DAT) transfected Chinese hamster ovary (CHO) cells, and dopamine reuptake blockade may provide an alternative approach for ameliorating parkinsonism. Here, we assessed the potential dopaminergic neuroprotective, and antiparkinsonian-like activity of BU. BU sample size was increased by using a scale-up extraction paradigm. Pharmacologically, BU significantly protected SK-N-SH cells from 1-methyl-4-phenylpyridinium (MPP⁺) insult, produced striking inhibitory actions on dopamine/norepinephrine uptake and WIN35,428 binding in synaptosomes on in vivo administration, and significantly preventing poor performance on rotarod and dopaminergic loss in substantia nigra in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) mice. BU acts by protecting dopaminergic neurons from MPP⁺ injury and preventing against MPTP-induced behavioral and histological lesions in the Parkinson’s disease (PD) model, possibly by inhibiting monoamine transporters. These findings suggest that BU could be meaningful in PD treatment. Received 14 January 2009; received after revision 22 February 2009; accepted 10 March 2009     

Glycoprotein IIb/IIIa blockade inhibits platelet aminophospholipid exposure by potentiating translocase and attenuating scramblase activity

The present study investigated the mechanisms underlying the inhibition of platelet phosphatidylserine (PS) exposure by GPIIb/IIIa blockade. Platelet PS exposure induced by thrombin stimulation was cell-cell contact dependent. GPIIb/IIIa blockade by c7E3 or SR121566 inhibited thrombin-induced platelet PS exposure. Thrombin stimulation induced mild, while A23187 induced extensive platelet-derived microparticle (PDMP) generation. Thrombin-induced PDMP generation was not inhibited by GPIIb/IIIa blockade. Aminophospholipid translocase activity was reduced upon platelet activation by thrombin. The reduction of non-PS-exposing platelets was attenuated by GPIIb/IIIa blockade, while little translocase activity was seen in PS-exposing platelets. Thrombin increased scramblase activity slightly in non-PS-exposing platelets, which was inhibited by GPIIb/IIIa blockade, and markedly enhanced scramblase activity in PS-exposing platelets. Activation of platelet calpain and caspase-3 or cytosolic calcium mobilization were not altered by GPIIb/IIIa inhibition. Thus, GPIIb/IIIa blockade inhibits platelet PS exposure by enhancing translocase activity and attenuating scramblase activity, but does not inhibit PDMP generation. Received 13 December 2006; received after revision 5 February 2007; accepted 9 March 2007     

The COMT inhibitor tolcapone potentiates the anticataleptic effect of Madopar in MPP+-lesioned mice

Orally administered Madopar (levodopa/benserazide 41) dose-dependently antagonized haloperidol-induced (1 mg/kg s.c.) catalepsy in MPP⁺-lesioned mice. Pretreatment with a new selective catechol-O-methyltransferase (COMT) inhibitor, tolcapone (30 mg/kg p.o.), slightly potentiated the antagonistic effect of Madopar (15 mg/kg p.o.) on haloperidol-induced catalepsy. The ability of tolcapone to increase the Madopar effect was significantly attenuated by high doses of 3-O-methyldopa (3-OMD) (800 mg/kg i.p.). This might suggest a competitive blockade of the active transport of levodopa through the blood-brain barrier. In conclusion, the inhibitory effect of tolcapone on the O-methylation of levodopa to 3-OMD by COMT is largely due to improved levodopa and dopamine availability in the brain, and to the reduced formation of 3-OMD.     

D-ala2-Metenkephalinamide blocks the synaptically elicited cortical spreading depression in rats

Summary Spreading depression (SD) was elicited in rats anesthetized with pentobarbital by a train of 8 electrical pulses (0.1 ms, 10 Hz) applied to parietal cortex. Local application of 50 g of D-ala²-metenkephalinamide (DAME) on the stimulated area evoked one or two SD waves followed by an increase of SD threshold from 40 V to 90 V. This effect could be partly prevented by naloxone (1 mg/kg i.p.) and reversed by local application of 4-aminopyridine (10^–3 M, 2 l), which reduced SD threshold to 5 and 20 V in normal and DAME-treated cortex, respectively. It is argued that DAME exerts an inhibitory effect on cortical neurons and that the initial SD facilitation is due to initial blockade of inhibitory neurons in the superficial cortical layers.supported by the European Training Program in Brain and Behavior Research.     

Naltrexone influence on hibernation

Summary In the garden dormouse, opiate receptor blockade by naltrexone decreased the score for sleeping behaviour during hibernation at 24.00 h, indicative of a possible involvement of endorphins in the control of hibernation.Acknowledgment. The author wishes to thank Dr B. Bondy, Dr J. Wissmann and Mr A. Wünsch for helpful assistance.     

Inhibition of SRC family kinases facilitates anti-CTLA4 immunotherapy in head and neck squamous cell carcinoma

The immune system plays a critical role in the establishment, development, and progression of head and neck squamous cell carcinoma (HNSCC). As treatment with single-immune checkpoint agent results in a lower response rate in patients, it is important to investigate new strategies to maintain favorable anti-tumor immune response. Herein, the combination immunotherapeutic value of CTLA4 blockade and SFKs inhibition was assessed in transgenic HNSCC mouse model. Our present work showed that tumor growth was not entirely controlled when HNSCC model mice were administered anti-CTLA4 chemotherapeutic treatment. Moreover, it was observed that Src family kinases (SFKs) were hyper-activated and lack of anti-tumor immune responses following anti-CTLA4 chemotherapeutic treatment. We hypothesized that activation of SFKs is a mechanism of anti-CTLA4 immunotherapy resistance. We, therefore, carried out combined drug therapy using anti-CTLA4 mAbs and an SFKs’ inhibitor, dasatinib. As expected, dasatinib and anti-CTLA4 synergistically inhibited tumor growth in Tgfbr1/Pten 2cKO mice. Furthermore, dasatinib and anti-CTLA4 combined to reduce the number of myeloid-derived suppressor cells and Tregs, increasing the CD8⁺ T cell-to-Tregs ratio. We also found that combining dasatinib with anti-CTLA4 therapy significantly attenuated the expression of p-STAT3^Y705 and Ki67 in tumoral environment. These results suggest that combination therapy with SFKs inhibitors may be a useful therapeutic approach to increase the efficacy of anti-CTLA4 immunotherapy in HNSCC.     

Different effects of the H2-antagonists on gastric emptying in the rat

Summary H₂-receptor antagonists, in doses capable of inhibiting gastric secretion, did not generally affect gastric emptying. Exceptions were burimamide, which delayed the emptying rate, and ranitidine, which accelerated it. At higher doses burimamide, metiamide cimetidine and oxmetidine delayed gastric emptying, but ranitidine accelerated it to a greater extent. Tiotidine remained ineffective. These data suggest that changes in emptying rate are independent of the H₂-receptor blockade.This work was supported by a grant from the C.N.R., Rome.     

Neuromuscular blocking activity of dibekacin,a new semisynthetic aminoglycoside antibiotic

Summary Dibekacin possesses the untoward effect of producing a neuromuscular blockade. Neostigmine is unable to reverse the neuromuscular blockade produced by dibekacin. Calcium has not only the ability to restore the neuromuscular transmission but also to exert protective action against the neuromuscular blocking activity of dibekacin.     

Negative inotropic effect of some H2-receptor antagonists on the isolated human atria

H2-Receptor antagonists were found to possess in various degrees a negative inotropic effect on human atria in vitro. This effect seemed to be independent of H2-receptor blockade and, at least in the case of oxmetidine, seemed to involve calcium ion transport and/or utilization.     

Is the renal vasodilatation induced byβ-adrenoceptor stimulants in the dog mediated through dopamine receptor?

Summary In chloralose-anaesthetized dogs the renal vasodilator effect of isoprenaline is depressed by blockade of either dopamine or -adrenoceptors but the renal vasodilator effect of dopamine is depressed only by blockade of dopamine receptors. This suggests that the vasodilatation induced by -stimulants within the canine kidney is due in part to activation of dopamine receptors.This study was supported by the Life Insurance Medical Research Fund of Australia and New Zealand and by the Australian Kidney Foundation.We thank Mr S. Marshall for technical assistance.     

Effect of adrenaline and adrenergic active drugs on growth hormone secretion in immature cockerels

Summary In immature cockerels adrenaline administration lowered the levels of plasma growth hormone. Both alpha and beta adrenergic receptor agonists also depressed the circulating growth hormone levels. In the presence of beta blockade, the suppressive effect of adrenaline on growth hormone secretion was not observed.     

Effect of adrenaline and adrenergic active drugs on growth hormone secretion in immature cockerels.

In immature cockerels adrenaline administration lowered the levels of plasma growth hormone. Both alpha and beta adrenergic receptor agonists also depressed the circulating growth hormone levels. In the presence of beta blockade, the suppressive effect of adrenaline on growth hormone secretion was not observed.     

Activation of sustained sympathetic vasodilatation in dog by spinal cord stimulation

Summary Electrical stimulation in lateral sites of the upper cervical spinal cord evoked vasodilatation after adrenergic blockade. Sympathetic fibres mediating sustained vasodilatation were shown to be separate from adrenergic sympathetic fibres since the adrenergic vasoconstrictor response in the paw evoked by vasomotor stimulation in the medulla was not reversed to vasodilatation after bretylium.Supported by USPHS Grant No. HL 08570 and part of thesis work by Dr.Rolewicz.     

Voltage-clamp analysis of the sodium and potassium currents in skeletal muscle fibres treated with 4-aminopyridine

Summary External application of low concentrations of 4-aminopyridine blocks potassium currents without affecting sodium currents in pieces of single frog skeletal muscle fibres. The blockade of potassium currents was voltage-dependent, being partially relieved on depolarization.This study was supported by a grant from the Oficina técnica de Desarrollo Cientifico of the University of Chile (Project 4437-R, 1977).     

Influence of ethanol on calcium,inositol phospholipids and intracellular signalling mechanisms

Summary Studies have implicated Ca⁺⁺ in the actions of ethanol at many biochemical levels. Calcium as a major intracellular messenger in the central nervous system is involved in many processes, including protein phosphorylation enzyme activation and secretion of hormones and neurotransmitters. The control of intracellular calcium, therefore, represents a major step by which neuronal cells regulate their activities. The present review focuses on three primary areas which influence intracellular calcium levels; voltage-dependent Ca⁺⁺ channels, receptor-mediated inositol phospholipid hydrolysis, and Ca⁺⁺/Mg⁺⁺-ATPase, the high affinity membrane Ca⁺⁺ pump.Current research suggests that a subtype of the voltage-dependent Ca⁺⁺ channel, the dihydropyridine-sensitive Ca⁺⁺ channel, is uniquely sensitive to acute and chronic ethanol treatment. Acute exposure inhibits, while chronic ethanol exposure increases⁴⁵Ca⁺⁺-influx and [³H]dihydropyridine receptor binding sites. In addition, acute and chronic exposure to ethanol inhibits, then increases Ca⁺⁺/Mg⁺⁺-ATPase activity in neuronal membranes. Changes in Ca⁺⁺ channel and Ca⁺⁺/Mg⁺⁺-ATPase activity following chronic ethanol may occur as an adaptation process to increase Ca⁺⁺ availability for intracellular processes. Since receptor-dependent inositol phospholipid hydrolysis is enhanced after chronic ethanol treatment, subsequent activation of protein kinase-C may also be involved in the adaptation process and may indicate increased coupling for receptor-dependent changes in Ca⁺⁺/Mg⁺⁺-ATPase activity.The increased sensitivity of three Ca⁺⁺-dependent processes suggest that adaptation to chronic ethanol exposure may involve coupling of one or more of these processes to receptor-mediated events.     

Influence of pyridinolcarbamate on hepatic cholesterogenesis in rats

Zusammenfassung Männliche Wistar-Ratten wurden während 3 Wochen mit 0,3% Pyridinolcarbamat gefüttert, der Cholesterinspiegel in Leber und Serum bestimmt und ausserdem der Einbau von Na-Acetat-1-¹⁴C und Mevalonsäure-2-¹⁴C im Cholesterin von Leberschnitten gemessen. Der Cholesterinspiegel in Serum und Leber wird durch Pyridinolcarbamat nicht beeinflusst. Der Einbau von Acetat-1-¹⁴C in Cholesterin wurde durch Pyridinolcarbamat gehemmt, während die Conversion von Mevalonat-2-¹⁴C unbeeinflusst blieb.